Prokaryotic expression of the thyrotropin receptor and identification of an immunogenic region of the protein using synthetic peptides

Osamu Takai, Rajesh K. Desai, Gattadahalli Seetharamaiah, Craig A. Jones, Graham P. Allaway, Takashi Akamizu, Leonard D. Kohn, Beller S. Prabhakar

Research output: Contribution to journalArticle

74 Citations (Scopus)

Abstract

Graves' disease is characterized by hypersecretion of thyroid hormones due to binding of autoantibodies to the thyrotropin receptor (TSHR). In order to study immunological aspects of the TSHR we expressed the extracellular domain of the rat TSHR (ETSHR) as a fusion protein with β-galactosidase in a prokaryotic system. The identity of this ETSHR-fusion protein was confirmed by Western blot, using antibodies to synthetic peptides derived from TSHR. Patients' sera reacted to a significantly greater extent with the affinity purified ETSHR relative to control sera. Similarly, sera from patients with Graves' disease displayed significant reactivity with only one of five peptides, RH2 (residues 352-366), when compared with normal sera. These data, together with the predicted hydrophilicity of the peptide RH2, suggest that amino acids 352-366 which lie within one of the unique regions of the extracellular domain of the TSHR may be important for antibody binding.

Original languageEnglish (US)
Pages (from-to)319-326
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume179
Issue number1
DOIs
StatePublished - Aug 30 1991
Externally publishedYes

Fingerprint

Thyrotropin Receptors
Peptides
Graves Disease
Fusion reactions
Serum
Galactosidases
Proteins
Antibodies
Hydrophilicity
Thyroid Hormones
Autoantibodies
Rats
Hydrophobic and Hydrophilic Interactions
Amino Acids
Western Blotting

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Prokaryotic expression of the thyrotropin receptor and identification of an immunogenic region of the protein using synthetic peptides. / Takai, Osamu; Desai, Rajesh K.; Seetharamaiah, Gattadahalli; Jones, Craig A.; Allaway, Graham P.; Akamizu, Takashi; Kohn, Leonard D.; Prabhakar, Beller S.

In: Biochemical and Biophysical Research Communications, Vol. 179, No. 1, 30.08.1991, p. 319-326.

Research output: Contribution to journalArticle

Takai, Osamu ; Desai, Rajesh K. ; Seetharamaiah, Gattadahalli ; Jones, Craig A. ; Allaway, Graham P. ; Akamizu, Takashi ; Kohn, Leonard D. ; Prabhakar, Beller S. / Prokaryotic expression of the thyrotropin receptor and identification of an immunogenic region of the protein using synthetic peptides. In: Biochemical and Biophysical Research Communications. 1991 ; Vol. 179, No. 1. pp. 319-326.
@article{37f15ef558db4452b27cbb05ce81542c,
title = "Prokaryotic expression of the thyrotropin receptor and identification of an immunogenic region of the protein using synthetic peptides",
abstract = "Graves' disease is characterized by hypersecretion of thyroid hormones due to binding of autoantibodies to the thyrotropin receptor (TSHR). In order to study immunological aspects of the TSHR we expressed the extracellular domain of the rat TSHR (ETSHR) as a fusion protein with β-galactosidase in a prokaryotic system. The identity of this ETSHR-fusion protein was confirmed by Western blot, using antibodies to synthetic peptides derived from TSHR. Patients' sera reacted to a significantly greater extent with the affinity purified ETSHR relative to control sera. Similarly, sera from patients with Graves' disease displayed significant reactivity with only one of five peptides, RH2 (residues 352-366), when compared with normal sera. These data, together with the predicted hydrophilicity of the peptide RH2, suggest that amino acids 352-366 which lie within one of the unique regions of the extracellular domain of the TSHR may be important for antibody binding.",
author = "Osamu Takai and Desai, {Rajesh K.} and Gattadahalli Seetharamaiah and Jones, {Craig A.} and Allaway, {Graham P.} and Takashi Akamizu and Kohn, {Leonard D.} and Prabhakar, {Beller S.}",
year = "1991",
month = "8",
day = "30",
doi = "10.1016/0006-291X(91)91372-J",
language = "English (US)",
volume = "179",
pages = "319--326",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Prokaryotic expression of the thyrotropin receptor and identification of an immunogenic region of the protein using synthetic peptides

AU - Takai, Osamu

AU - Desai, Rajesh K.

AU - Seetharamaiah, Gattadahalli

AU - Jones, Craig A.

AU - Allaway, Graham P.

AU - Akamizu, Takashi

AU - Kohn, Leonard D.

AU - Prabhakar, Beller S.

PY - 1991/8/30

Y1 - 1991/8/30

N2 - Graves' disease is characterized by hypersecretion of thyroid hormones due to binding of autoantibodies to the thyrotropin receptor (TSHR). In order to study immunological aspects of the TSHR we expressed the extracellular domain of the rat TSHR (ETSHR) as a fusion protein with β-galactosidase in a prokaryotic system. The identity of this ETSHR-fusion protein was confirmed by Western blot, using antibodies to synthetic peptides derived from TSHR. Patients' sera reacted to a significantly greater extent with the affinity purified ETSHR relative to control sera. Similarly, sera from patients with Graves' disease displayed significant reactivity with only one of five peptides, RH2 (residues 352-366), when compared with normal sera. These data, together with the predicted hydrophilicity of the peptide RH2, suggest that amino acids 352-366 which lie within one of the unique regions of the extracellular domain of the TSHR may be important for antibody binding.

AB - Graves' disease is characterized by hypersecretion of thyroid hormones due to binding of autoantibodies to the thyrotropin receptor (TSHR). In order to study immunological aspects of the TSHR we expressed the extracellular domain of the rat TSHR (ETSHR) as a fusion protein with β-galactosidase in a prokaryotic system. The identity of this ETSHR-fusion protein was confirmed by Western blot, using antibodies to synthetic peptides derived from TSHR. Patients' sera reacted to a significantly greater extent with the affinity purified ETSHR relative to control sera. Similarly, sera from patients with Graves' disease displayed significant reactivity with only one of five peptides, RH2 (residues 352-366), when compared with normal sera. These data, together with the predicted hydrophilicity of the peptide RH2, suggest that amino acids 352-366 which lie within one of the unique regions of the extracellular domain of the TSHR may be important for antibody binding.

UR - http://www.scopus.com/inward/record.url?scp=0025939913&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025939913&partnerID=8YFLogxK

U2 - 10.1016/0006-291X(91)91372-J

DO - 10.1016/0006-291X(91)91372-J

M3 - Article

VL - 179

SP - 319

EP - 326

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 1

ER -